Glucagon-like peptide1 receptor agonists

ABSTRACT

In an embodiment, the present invention provides a compound of the formula:or a pharmaceutically acceptable salt thereof, and methods of using this compound for treating type II diabetes mellitus.

This invention relates to glucagon-like peptide-1 receptor agonists andtherapeutic uses of the compounds to treat type II diabetes mellitus.

Glucagon-like peptide-1 (GLP-1) is a member of the incretin family ofpeptide hormones secreted by intestinal enteroendocrine L-cells. GLP-1induces the release of insulin from beta cells in a glucose dependentmanner. However, GLP-1 is rapidly metabolized so that only a smallpercentage of the GLP-1 can be utilized to induce insulin secretion. Tooffset this, GLP-1 receptor (GLP-1R) agonists have been developed toenhance insulin secretion as a treatment for type II diabetes mellitus.

The majority of GLP-1R agonists that have been approved to treat type IIdiabetes mellitus are injectable agents. Patients often prefer orallyadministered drugs because of the drawbacks associated with injectionsuch as inconvenience, pain, and the potential for injection siteirritation.

WO2018/109607 discloses certain benzimidazole derivatives, which aredescribed as GLP-1R agonists.

However, there is a need for alternative GLP-1R agonists. In particular,there is a need for GLP-1R agonists which can be administered orally.There is especially a need for GLP-1R agonists having improved potency,a favourable toxicology profile and/or a pharmacokinetic profile whichsupports once daily dosing.

Accordingly, the present invention provides a compound of the formula:

wherein

R¹ is H or F; R² is H or F; and

R³ is H or CH₃;or a pharmaceutically acceptable salt thereof.

Formula I includes all individual enantiomers, and mixtures thereof, aswell as racemates, and pharmaceutically acceptable salts thereof.

In an embodiment, there is a provided a compound of the formula:

or a pharmaceutically acceptable salt thereof.

In an embodiment, there is provided a compound of the formula:

wherein R¹ is H or F, or a pharmaceutically acceptable salt thereof.

In an embodiment, there is provided a compound of the formula:

wherein R¹ is H or F, or a pharmaceutically acceptable salt thereof.

In one embodiment, the compound is a compound of the formula:

or a pharmaceutically acceptable salt thereof. In a preferredembodiment, the compound is a compound of the formula:

or a pharmaceutically acceptable salt thereof.

In one embodiment, the compound is a compound of the formula:

or a pharmaceutically acceptable salt thereof. In a preferredembodiment, the compound is a compound of the formula:

or a pharmaceutically acceptable salt thereof. In a particularlypreferred embodiment, there is provided the tert-butylamine salt (alsoknown as the erbumine salt) of:

In an embodiment, there is provided a compound of the formula:

wherein R² is H or F, or a pharmaceutically acceptable salt thereof.

In an embodiment, there is provided a compound of the formula:

wherein R² is H or F, or a pharmaceutically acceptable salt thereof.

In one embodiment, the compound is a compound of the formula:

or a pharmaceutically acceptable salt thereof. In a preferredembodiment, the compound is a compound of the formula:

or a pharmaceutically acceptable salt thereof.

In one embodiment, the compound is a compound of the formula:

or a pharmaceutically acceptable salt thereof. In a preferredembodiment, the compound is a compound of the formula:

or a pharmaceutically acceptable salt thereof.

Formula I encompasses Formulae Ia, Ib, II, IIa, IIb, III, Ma and Mb andreference to Formula I below, for example in the methods of treatmentand therapeutic uses, is also to be read as a reference to each and allof these sub-formulae.

In another embodiment, there is provided a pharmaceutically acceptablecomposition comprising a compound of Formula I, or a pharmaceuticallyacceptable salt thereof, and at least one of a pharmaceuticallyacceptable carrier, diluent or excipient. In a preferred embodiment, thepharmaceutically acceptable composition is formulated for oraladministration.

In another embodiment, there is provided a method of treating a mammalfor type II diabetes mellitus, the method comprises administering to themammal in need of treatment a pharmaceutically acceptable compositioncomprising an effective amount of a compound of Formula I, or apharmaceutically acceptable salt thereof, and at least one of apharmaceutically acceptable carrier, diluent or excipient. In oneembodiment, the pharmaceutically acceptable composition is formulatedfor oral administration. Preferably, the mammal is a human.

In another embodiment, there is provided a method of treating a mammalfor type II diabetes mellitus, the method comprises administering to themammal in need of treatment an effective amount of a compound of FormulaI, or a pharmaceutically acceptable salt thereof. In a preferredembodiment, the mammal is a human.

In another embodiment, there is provided a method of lowering bloodglucose levels in a mammal, the method comprises administering to themammal in need of treatment an effective amount of a compound of FormulaI, or a pharmaceutically acceptable salt thereof. In a preferredembodiment, the mammal is a human.

In another embodiment, there is provided a method of treatinghyperglycemia in a mammal, the method comprises administering to themammal in need of treatment an effective amount of a compound of FormulaI, or a pharmaceutically acceptable salt thereof. In a preferredembodiment, the mammal is a human.

In an embodiment, there is provided a compound of Formula I, or apharmaceutically acceptable salt thereof, for use in therapy.

In another embodiment, there is provided a compound of Formula I, or apharmaceutically acceptable salt thereof, for use in the treatment oftype II diabetes mellitus.

In another embodiment, there is provided a compound of Formula I, or apharmaceutically acceptable salt thereof, for use in lowering bloodglucose levels.

In another embodiment, there is also provided a compound of Formula I,or a pharmaceutically acceptable salt thereof, for use in treatinghyperglycemia.

In an embodiment, there is provided the use of a compound of Formula I,or a pharmaceutically acceptable salt thereof, for the manufacture of amedicament for the treatment of type II diabetes mellitus.

In an embodiment, there is provided the use of a compound of Formula I,or a pharmaceutically acceptable salt thereof, for the manufacture of amedicament for lowering blood glucose levels.

In an embodiment, there is provided the use of a compound of Formula I,or a pharmaceutically acceptable salt thereof, for the manufacture of amedicament for the treatment of hyperglycemia.

In a preferred embodiment, the compound of Formula I is administeredorally. In a preferred embodiment, the compound of Formula I isadministered once daily. In another preferred embodiment, thetherapeutic use is in a human.

The term “pharmaceutically acceptable salt” as used herein refers a saltof a compound of the invention considered to be acceptable for clinicaland/or veterinary use. Examples of pharmaceutically acceptable salts andcommon methodologies for preparing them can be found in “Handbook ofPharmaceutical Salts: Properties, Selection and Use” P. Stahl, et al.,2nd Revised Edition, Wiley-VCH, 2011 and S. M. Berge, et al.,“Pharmaceutical Salts”, Journal of Pharmaceutical Sciences, 1977, 66(1),1-19.

Examples of pharmaceutical compositions and processes for theirpreparation can be found in “Remington: The Science and Practice ofPharmacy”, Loyd, V., et al. Eds., 22^(nd) Ed., Mack Publishing Co.,2012. In one embodiment, the pharmaceutically compositions can beformulated for oral administration. Preferably the pharmaceuticalcompositions are formulated as a tablet, capsule, or a solution. Thetablet, capsule, or solution can include a compound of Formula I in anamount effective for treating a patient in need of treatment.

The term “effective amount” refers to the amount or dose of a compoundof Formula I, or a pharmaceutically acceptable salt thereof, which, uponsingle or multiple dose administration to the patient, provides thedesired effect in the patient under diagnosis or treatment. Theattending physician, as one skilled in the art, can readily determine aneffective amount by the use of conventional techniques and by observingresults obtained under analogous circumstances. Factors considered inthe determination of an effective amount or dose of a compound include:whether the compound or its salt will be administered; theco-administration of other agents, if used; the species of mammal to betreated; its size, age, and general health; the degree of involvement orthe severity of the disorder; the response of the individual mammal; themode of administration; the bioavailability characteristics of thepreparation administered; the dose regimen selected; and other relevantcircumstances. The compounds of the present invention are effective at adosage per day that falls within the range of about 0.01 to about 15mg/kg of body weight.

As used herein, the terms “treating”, “to treat”, or “treatment”, refersto lowering, reducing, or reversing the progression or severity of anexisting symptom, disorder, or condition, such as hyperglycemia, whichcan include increasing insulin secretion.

The compounds of Formula I can be formulated as pharmaceuticalcompositions administered by any route which makes the compoundbioavailable. Preferably, such compositions are for oral administration.Such pharmaceutical compositions and processes for preparing same arewell known in the art (See, e.g., Remington, J. P., “Remington: TheScience and Practice of Pharmacy”, L. V. Allen, Editor, 22^(nd) Edition,Pharmaceutical Press, 2012).

The compounds of Formula I and the pharmaceutically acceptable saltsthereof are useful in the therapeutic uses of the invention, withcertain configurations being preferred.

Compounds of the present invention include:

or pharmaceutically acceptable salts thereof.

Further compounds of the present invention include:

or pharmaceutically acceptable salts thereof.

Further compounds of the present invention include:

or pharmaceutically acceptable salts thereof.

Although the present invention contemplates all individual enantiomers,mixtures thereof, and racemates, compounds of Formula Ia, IIa and IIIa,and pharmaceutically acceptable salts thereof, are particularlypreferred.

Individual enantiomers may be separated or resolved by one of ordinaryskill in the art at any convenient point in the synthesis of compoundsof the invention, by methods such as selective crystallizationtechniques, chiral chromatography (See for example, J. Jacques, et al.,“Enantiomers, Racemates, and Resolutions”, John Wiley and Sons, Inc.,1981, and E. L. Eliel and S. H. Wilen, “Stereochemistry of OrganicCompounds”, Wiley-Interscience, 1994), or supercritical fluidchromatography (SFC) (See for example, T. A. Berger; “SupercriticalFluid Chromatography Primer,” Agilent Technologies, July 2015).

A pharmaceutically acceptable salt of the compounds of the invention canbe formed, for example, by reaction of a compound of Formula I and anappropriate pharmaceutically acceptable base in a suitable solvent understandard conditions well known in the art (See, for example, Bastin, R.J., et al.; Org. Process. Res. Dev., 4, 427-435, 2000 and Berge, S. M.,et al.; J. Pharm. Sci., 66, 1-19, 1977). A preferred salt is thetert-butyl amine (or erbumine) salt.

Certain abbreviations used herein are defined according to Daub G. H.,et al., “The Use of Acronyms in Organic Chemistry” Aldrichimica Acta,1984, 17(1), 6-23. Certain abbreviations are defined as follows: “ACN”refers to acetonitrile; “ATP” refers to adenosine triphosphate; “BSA”refers to Bovine Serum Albumin; “cAMP” refers to cyclicadenosine-3′,5′-monophosphate; “DCM” refers to dichloromethane ormethylene chloride; “DIPEA” refers to N,N-diisopropylethylamine; “DMF”refers to N,N-dimethylformamide; “DMSO” refers to dimethyl sulfoxide;“EC₅₀” refers to the concentration of an agent which produces 50%response of the target activity compared to a predefined positivecontrol compound (absolute EC₅₀); “ES/MS” refers to electrospray massspectrometry; “EtOAc” refers to ethyl acetate; “HATU” refers to1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium3-oxid hexafluorophosphate; “HEK” refers to human embryonic kidney;“HEPES” refers to 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid;“h” refers to hours or hour, respectively; “MeOH” refers to methanol ormethyl alcohol; “min” refers to minute or minutes; “Pd(dppf)Cl₂” refersto [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II); “RT”refers to room temperature; and “THF” refers to tetrahydrofuran.

The compounds of the present invention may be prepared by a variety ofprocedures, some of which are illustrated in the Preparations andExamples below. The specific synthetic steps for each of the routesdescribed may be combined in different ways, to prepare compounds of theinvention, or salts thereof. The product of each step below can berecovered by conventional methods, including extraction, evaporation,precipitation, chromatography, filtration, trituration, andcrystallization. The reagents and starting materials are readilyavailable to one of ordinary skill in the art. Individual isomers,enantiomers, and diastereomers may be separated or resolved at anyconvenient point in the synthesis, by methods such as, selectivecrystallization techniques or chiral chromatography (See for example, J.Jacques, et al., “Enantiomers, Racemates, and Resolutions”, John Wileyand Sons, Inc., 1981, and E. L. Eliel and S. H. Wilen, “Stereochemistryof Organic Compounds”, Wiley-Interscience, 1994). Without limiting thescope of the invention, the following preparations, and examples areprovided to further illustrate the invention.

Scheme 1 shows the synthesis of intermediate 6, which is used in thepreparation of the compounds of Formula I. Benzoic acid 1 firstundergoes reduction with borane dimethylsulfide complex in Step 1 togive alcohol 2. The alcohol is converted into a leaving group (LG,intermediate 3). For example, the alcohol in intermediate 2 can beconverted to a mesylate group using methanesulfonyl chloride at −15° C.in Step 2, or it can be converted to a bromide using phosphorustribromide at 0° C. Intermediate 3 is reacted with NaCN in Step 3 togive nitrile 4. Nitrile 4 is converted with KOH at elevated temperaturein Step 4 to give acid 5, which is then esterified in Step 5 to giveintermediate 6 using oxalyl chloride, DMF, and methanol.

Scheme 2 depicts the preparation of key intermediate 12 for thepreparation of the compounds of Formula I via two routes. In the firstroute, aryl halide 6 undergoes a one-pot Miyura borylation/Suzukicoupling: using bis(pinacolato)diboron, Pd(dppf)Cl₂, and potassiumacetate at elevated temperature, aryl halide 6 is converted in Step 1ato boronic ester 7, whereupon bromopyridine 8 and K₂CO₃ are added to thereaction (Step 2a) giving intermediate 12. In the second route, atwo-step process is employed: Suzuki coupling of aryl halide 6 with6-hydroxypyridine-2-boronic acid pinacol ester 9 using Pd(dppf)Cl₂ andK₂CO₃ at elevated temperature (Step 1b) provides intermediate 10, whichis then alkylated with 4-(bromomethyl)-3-fluorobenzonitrile 6 usingAg₂CO₃ at elevated temperature (Step 2b) to give intermediate 12. Esterhydrolysis of intermediate 12 in Step 3 using LiOH yields acidintermediate 13.

Alternatively, key intermediates 12 and 13 can be prepared according toScheme 3, coupling bromopyridine 8 with boronic ester 7 or boronic acid14 using Pd(dppf)Cl₂ and potassium carbonate at elevated temperature.

Scheme 4 shows the conversion of key intermediate 13 to compounds ofFormula I. Amide coupling in Step 1 using HATU and dianiline 15 givesintermediate 16. Cyclization (Step 2) is accomplished by heatingintermediate 16 in acetic acid to give benzimidazole 17. Finally, inStep 3 the compounds of Formula I are obtained by hydrolysis of 17 usingLiOH.

PREPARATIONS AND EXAMPLES

LC-ES/MS is performed on an AGILENT® HP1200 liquid chromatographysystem. Electrospray mass spectrometry measurements (acquired inpositive and/or negative mode) are performed on a Mass SelectiveDetector quadrupole mass spectrometer interfaced to an HPLC which may ormay not have an ELSD. LC-ES/MS conditions (low pH): column: PHENOMENEX®GEMINI® NX C18 2.0×50 mm 3.0 μm, 110 Å; gradient: 5-95% B in 1.5 min,then 95% B for 0.5 min column temperature: 50° C.+/−10° C.; flow rate:1.2 mL/min; 1 μL injection volume; Solvent A: deionized water with 0.1%HCOOH; Solvent B: ACN with 0.1% formic acid; wavelength 200-400 nm and212-216 nm. If the HPLC is equipped with an ELSD the settings are 45° C.evaporator temperature, 40° C. nebulizer temperature, and 1.6 SLM gasflow rate. Alternate LC-MS conditions (high pH): column: Waters xBridge®C18 column 2.1×50 mm, 3.5 μm; gradient: 5-95% B in 1.5 min, then 95% Bfor 0.50 min; column temperature: 50° C.+/−10° C.; flow rate: 1.2mL/min; 1 μL injection volume; Solvent A: 10 mM NH₄HCO₃ pH 9; Solvent B:ACN; wavelength: 200-400 nm and 212-216 nm; if had ELSD: 45° C.evaporator temp, 40° C. nebulizer temp, and 1.60 SLM gas flow rate.

The X-ray powder diffraction (XRPD) patterns of crystalline solids areobtained on a Bruker D4 Endeavor X-ray powder diffractometer, equippedwith a CuKα source and a Vantec detector, operating at 35 kV and 50 mA.The sample is scanned between 4 and 40 2θ°, with a step size of 0.0082θ° and a scan rate of 0.5 seconds/step, and using 1.0 mm divergence,6.6 mm fixed anti-scatter, and 11.3 mm detector slits. The dry powder ispacked on a quartz sample holder and a smooth surface is obtained usinga glass slide. The crystal form diffraction patterns are collected atambient temperature and relative humidity. Crystal peak positions aredetermined in MDI-Jade after whole pattern shifting based on an internalNIST 675 standard with peaks at 8.853 and 26.774 2θ°. It is well knownin the crystallography art that, for any given crystal form, therelative intensities of the diffraction peaks may vary due to preferredorientation resulting from factors such as crystal morphology and habit.Where the effects of preferred orientation are present, peak intensitiesare altered, but the characteristic peak positions of the polymorph areunchanged. See, e.g. The United States Pharmacopeia #23, NationalFormulary #18, pages 1843-1844, 1995. Furthermore, it is also well knownin the crystallography art that for any given crystal form the angularpeak positions may vary slightly. For example, peak positions can shiftdue to a variation in the temperature at which a sample is analyzed,sample displacement, or the presence or absence of an internal standard.In the present case, a peak position variability of ±0.2 2θ° is presumedto take into account these potential variations without hindering theunequivocal identification of the indicated crystal form. Confirmationof a crystal form may be made based on any unique combination ofdistinguishing peaks.

Preparation 1 (4-Bromo-2-fluoro-5-methylphenyl)methanol

To a flask add: 4-bromo-2-fluoro-5-methylbenzoic acid (100 g, 421 mmol),THF (200 mL) and borane (dimethyl sulfide complex, 2 mol/L solution inTHF, 210 mL, 10 mmol). Stir the mixture at RT overnight. Quench thereaction mixture with HCl (1.0 N aqueous solution, 50 mL) and filter themixture. Concentrate the filtrate in-vacuo and partition the residuebetween EtOAc (400 mL) and water (400 mL). Wash the organics withsaturated aqueous NaCl (400 mL), dry over Na₂SO₄, filter, andconcentrate to give the title compound as solid (93.5 g, 99%). ¹H-NMR(400 MHz, CDCl₃) δ 7.29 (d, J=7.9 Hz, 1H), 7.26 (d, J=9.1 Hz, 1H), 4.69(s, 2H), 2.38 (s, 3H).

Preparation 2 (4-Bromo-2-fluoro-3-methyl-phenyl)methanol

Prepare the title compound essentially as described in Preparation 1using 4-bromo-2-fluoro-3-methylbenzoic acid. Purify the product bysilica gel chromatography using a gradient of 10 to 35% EtOAc inhexanes. LC-ES/MS peak retention time: 1.01 min.

Preparation 3 2-(4-Bromo-2-fluoro-5-methylphenyl)acetonitrile

Dissolve (4-bromo-2-fluoro-5-methylphenyl)methanol (92 g, 420 mmol) inDCM (500 mL) and add triethylamine (120 mL, 861 mmol). Cool the mixtureto −15° C. and add a solution of methanesulfonyl chloride (40 mL, 517mmol) in DCM (30 mL) dropwise to the reaction mixture. Stir the mixturefor 30 min at RT. Partition the reaction mixture between DCM (500 mL)and water (500 mL). Wash the organics with saturated aqueous NaCl (500mL), dry over Na₂SO₄, filter, and concentrate. Dissolve the residue inDMF (400 mL) and cool the mixture with an ice bath. Add NaCN (21.0 g,429 mmol) in one portion to the reaction mixture and stir at RTovernight. Partition the mixture between EtOAc (400 mL) and water (500mL). Wash the organics with saturated aqueous NaCl (500 mL), dry overNa₂SO₄, filter, and concentrate. Purify the residue by silica gelchromatography using a gradient of 10 to 30% EtOAc in hexanes to givethe title compound (47.0 g, 48%) as an oil. ¹H-NMR (400 MHz, CDCl₃) δ7.34 (d, J=8.7 Hz, 1H), 7.32 (d, J=8.1 Hz, 1H), 3.71 (s, 2H), 2.41 (s,3H).

Preparation 4 2-(4-Bromo-2-fluoro-3-methyl-phenyl)acetonitrile

Mix together (4-bromo-2-fluoro-3-methyl-phenyl)methanol (1.90 g, 8.67mmol) and DCM (20 mL). Cool the mixture to 0° C., then add phosphorustribromide (1.0 mL, 11 mmol) dropwise. Stir the mixture at 0° C. for 15min, then basify the mixture with saturated aqueous NaHCO₃ (10 mL).Extract the mixture with DCM (40 mL). Wash the organics with brine (30mL), dry over (Na₂SO₄), filter and concentrate to give a solid. Dissolvethe solid in DMSO (10 mL), then add NaCN (0.60 g, 13.0 mmol) and stirfor 1 h. Partition the mixture between EtOAc (50 mL) and water (50 mL).Wash the organics with brine (50 mL), dry over Na₂SO₄, filter andconcentrate to give the product as solid (1.3 g, 64%). LC-ES/MS peakretention time: 1.17 min

Preparation 5 Methyl 2-(4-bromo-2-fluoro-5-methyl-phenyl)acetate

To a flask add: 2-(4-bromo-2-fluoro-5-methylphenyl)acetonitrile (1.20 g,5.10 mmol), ethanol (5 mL), water (3 mL), and potassium hydroxide (0.90g, 16 mmol). Heat the mixture at 90° C. overnight. Cool the mixture withan ice bath and acidify with 1.0 M HCl to pH 4-5, then partition themixture between EtOAc (30 mL) and water (30 mL). Wash the organics withsaturated aqueous NaCl (30 mL), dry over Na₂SO₄, filter, and concentrateto give 2-(4-bromo-2-fluoro-5-methyl-phenyl)acetic acid as solid.Dissolve this in DCM (10 mL), then add DMF (0.05 mL, 0.6 mmol) andoxalyl chloride (0.5 mL, 6 mmol) at RT. Stir the mixture RT for 30 min,then add MeOH (2 mL, 49.4 mmol) dropwise. After 30 min, remove thesolvent in-vacuo and partition the residue between EtOAc (40 mL) and 5%NaHCO₃ (30 mL). Wash the organics with saturated aqueous NaCl (40 mL),dry over Na₂SO₄, filter, and concentrate to give the title compound asan oil (1.1 g, 80%). ES/MS m/z (⁷⁹Br,⁸¹Br) 278,280 (M+NH₄ ⁺).

Preparation 6 Methyl 2-(4-bromo-2-fluoro-3-methyl-phenyl)acetate

Prepare the title compound essentially as described in Preparation 5using 2-(4-bromo-2-fluoro-3-methyl-phenyl)acetonitrile. LC-ES/MS peakretention time: 1.22 min.

Preparation 7 Methyl 2-(4-bromo-2,6-difluorophenyl)acetate

Mix 4-bromo-2,6-difluorophenylacetic acid (3.30 g, 12.5 mmol), DCM (20mL), DMF (0.05 mL, 0.6 mmol), and oxalyl chloride (1.3 mL, 15 mmol).Stir the mixture at RT for 30 min, then add MeOH (1.5 mL, 37 mmol, 100mass %) dropwise. Concentrate the mixture and partition between EtOAc(30 mL) and saturated aqueous NaHCO₃ (15 mL). Wash the organics withsaturated aqueous NaCl (30 mL), dry over Na₂SO₄, filter, and concentrateto give the title compound as an oil (3.41 g, quantitative yield), whichis used without further purification in Preparation 10. ES/MS m/z(⁷⁹Br,⁸¹Br) 265,267 (M+H).

Preparation 8 4-[(6-Bromo-2-pyridyl)oxymethyl]-3-fluoro-benzonitrile

Dissolve 2-bromo-6-fluoropyridine (2.50 g, 13.8 mmol) and3-fluoro-4-(hydroxymethyl)benzonitrile (2.15 g, 13.8 mmol) in1,4-dioxane (25 mL) and add a solution of potassium tert-butoxide (20 wt% in THF, 10.0 mL, 16.6 mmol) dropwise over 12 min at RT. Heat thereaction mixture at 40° C. for 30 min. Pour the mixture into aqueousK₂CO₃ (1M) and extract twice with EtOAc. Wash the organics with waterand saturated aqueous NaCl, dry over Na₂SO₄, filter, and concentrate.Dry the residue in a vacuum oven at 50° C. to give the title compound(4.23 g, 95%) as a light yellow solid. ES/MS m/z (⁷⁹Br,⁸¹Br) 307,309(M+H).

Preparation 9 Methyl2-[2-fluoro-4-(6-hydroxy-2-pyridyl)-5-methyl-phenyl]acetate

To a flask add 6-hydroxypyridine-2-boronic acid pinacol ester (1.6 g,6.9 mmol), methyl 2-(4-bromo-2-fluoro-5-methyl-phenyl)acetate (2.2 g,8.4 mmol), THF (15 mL), water (1 mL), and potassium carbonate (2.0 g, 14mmol). Purge the mixture with nitrogen for 10 min, then add Pd(dppf)Cl₂(0.26 g, 0.35 mmol) and heat at 75° C. for 2 h. Partition the mixturebetween EtOAc (30 mL) and water (30 mL). Wash the organics withsaturated aqueous NaCl (30 mL), dry over Na₂SO₄, filter and concentrateto give the title compound (1.4 g, 74%) as a solid. ES/MS m/z 276 (M+H),274 (M−H).

Preparation 10 Methyl2-[2,6-difluoro-4-(6-hydroxy-2-pyridyl)phenyl]acetate

Prepare the title compound essentially as described in Preparation 9using methyl 2-(4-bromo-2,6-difluorophenyl)acetate, heating the reactionat 75° C. overnight. ES/MS m/z 280 (M+H).

Preparation 11 Methyl2-[2-fluoro-4-(6-hydroxy-2-pyridyl)-3-methyl-phenyl]acetate

Prepare the title compound essentially as described in Preparation 9using methyl 2-(4-bromo-2-fluoro-3-methyl-phenyl)acetate, heating thereaction at 75° C. overnight (18 h). ES/MS m/z 276 (M+H), 274 (M−H).

Preparation 12 Methyl2-[4-[6-[(4-cyano-2-fluoro-phenyl)methoxy]-2-pyridyl]-3-methyl-phenyl]acetate

Dissolve 2-(4-bromo-3-methylphenyl)acetic acid (10.7 g, 45.8 mmol) inDCM (50 mL). Cool the mixture in an ice/water bath, and then add oxalylchloride (4.8 mL, 55 mmol) and DMF (0.1 mL). Remove ice/water bath andstirred at RT for 2 h. Add MeOH (6.0 mL) dropwise over 2 min and stirredat RT for 1 h. Concentrate the reaction mixture in-vacuo and dissolvethe residue in EtOAc. Wash the organics with saturated aqueous NaHCO₃and saturated aqueous NaCl. Dry the organics over Na₂SO₄, then filterand concentrate. To the residue add bis(pinacolato)diboron (12.8 g, 50.4mmol) and potassium acetate (13.6 g, 137 mmol). Bubble nitrogen throughthe reaction mixture for 15 min, then add Pd(dppf)Cl₂ (complex with DCM,1.13 g, 1.37 mmol). Heat the reaction under nitrogen at 85° C. for 15 hin an oil bath, then remove the reaction flask from the oil bath.Dissolve potassium carbonate (9.49 g, 68.7 mmol) in water (60 mL),bubble nitrogen through the solution for 10 min, and then add thissolution to the reaction mixture followed by4-[(6-bromo-2-pyridyl)oxymethyl]-3-fluoro-benzonitrile (14.1 g, 45.8mmol). Bubble nitrogen through the entire reaction mixture for 5 min andheat under nitrogen at 85° C. for 6 h. Cool the reaction to near RT andconcentrate in-vacuo to remove most of the 1,4-dioxane. Dilute thismixture with EtOAc (200 mL) and wash with water and saturated aqueousNaCl. Dry the organics over Na₂SO₄, then filter and concentrate. Purifythe crude product by silica gel chromatography using a gradient of 5 to50% EtOAc in hexanes to give the title compound (13.3 g, 70%) as a lightyellow solid. ES/MS m/z 391 (M+H).

Preparation 13 Methyl2-[4-[6-[(4-cyano-2-fluoro-phenyl)methoxy]-2-pyridyl]-2-fluoro-5-methyl-phenyl]acetate

To a flask add methyl2-[2-fluoro-4-(6-hydroxy-2-pyridyl)-5-methyl-phenyl]acetate (1.40 g,5.09 mmol), 1,4-dioxane (35 mL), silver carbonate (1.7 g, 6.2 mmol), and4-(bromomethyl)-3-fluorobenzonitrile (1.4 g, 6.2 mmol). Heat the mixtureat 60° C. overnight. Filter off the solid and concentrate the filtrate.Purified the residue by silica gel chromatography using 12 to 55% EtOAcin hexanes to give the title compound (1.60 g, 77%) as a solid. ES/MSm/z 409 (M+H), 407 (M−H).

Preparation 14 Methyl2-[4-[6-[(4-cyano-2-fluoro-phenyl)methoxy]-2-pyridyl]-2-fluoro-phenyl]acetate

Charge a flask with4-[(6-bromo-2-pyridyl)oxymethyl]-3-fluoro-benzonitrile (2.02 g, 6.58mmol), methyl2-(2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)acetate(2.99 g, 9.88 mmol), K₂CO₃ (2.30 g, 16.5 mmol), 1,4-dioxane (30 mL) andwater (10 mL). Bubble nitrogen through the mixture for 10 min. AddPd(dppf)Cl₂ DCM complex (492 mg, 0.658 mmol) to the mixture and heat to80° C. under nitrogen for 5 h. Cool the reaction mixture, dilute withEtOAc (75 mL) and filter through a pad of Celite®. Wash the filtratewith water and saturated aqueous NaCl, dried over Na₂SO₄, filtered andconcentrated. Purify the resulting residue by silica gel chromatographywith a gradient of 5 to 90% EtOAc in hexanes to obtain the titlecompound (2.68 g, 94%). ES/MS m/z 395 (M+H).

Preparation 15 Methyl2-[4-[6-[(4-cyano-2-fluoro-phenyl)methoxy]-2-pyridyl]-2,6-difluoro-phenyl]acetate

Prepare the title compound essentially as described in Preparation 13using methyl 2-[2,6-difluoro-4-(6-hydroxy-2-pyridyl)phenyl]acetate,heating the reaction at 80° C. overnight. ES/MS m/z 413 (M+H).

Preparation 16 Methyl2-[4-[6-[(4-cyano-2-fluoro-phenyl)methoxy]-2-pyridyl]-2-fluoro-3-methyl-phenyl]acetate

Prepare the title compound essentially as described in Preparation 13using methyl2-[2-fluoro-4-(6-hydroxy-2-pyridyl)-3-methyl-phenyl]acetate, heating thereaction at 80 ° C. for 3 h. ES/MS m/z 409 (M+H).

Preparation 172-[4-[6-[(4-Cyano-2-fluoro-phenyl)methoxy]-2-pyridyl]-3-methyl-phenyl]aceticacid

To a flask add methyl2-[4-[6-[(4-cyano-2-fluoro-phenyl)methoxy]-2-pyridyl]-3-methyl-phenyl]acetate(1.20 g, 3.07 mmol), ACN (20 mL), water (10 mL), and lithium hydroxide(0.35 g, 15 mmol). Heat the mixture at 45° C. for 3 h. Cool the mixturewith an ice bath and acidify with 1.0 M HCl to pH=4-5. Partition themixture between EtOAc (30 mL) and water (30 mL). Wash the organics withbrine (30 mL), dry over Na₂SO₄, filter, and concentrate to give thetitle compound (1.1 g, 95%) as solid. ES/MS m/z 377 (M+H).

Preparation 182-[4-[6-[(4-Cyano-2-fluoro-phenyl)methoxy]-2-pyridyl]-2-fluoro-5-methyl-phenyl]aceticacid

To a vial add methyl2-[4-[6-[(4-cyano-2-fluoro-phenyl)methoxy]-2-pyridyl]-2-fluoro-5-methyl-phenyl]acetate(1.6 g, 3.9 mmol), ACN (20 mL), water (6 mL), and lithium hydroxide(0.45 g, 19 mmol). Heat the mixture at 45° C. for 2 h, cool the mixturewith an ice bath, and acidify with 1.0 M HCl to pH=4-5. Partition themixture between EtOAc (50 mL) and water (50 mL). Wash the organics withsaturated aqueous NaCl (50 mL), dry over Na₂SO₄, filter and concentratedto give the title compound (1.55 g, 100%) as solid. ES/MS m/z 395 (M+H).

Preparation 192-[4-[6-[(4-Cyano-2-fluoro-phenyl)methoxy]-2-pyridyl]-2-fluoro-phenyl]aceticacid

Dissolve methyl2-[4-[6-[(4-cyano-2-fluoro-phenyl)methoxy]-2-pyridyl]-2-fluoro-phenyl]acetate(2.68 g, 6.25 mmol) in THF (50 mL), then add lithium hydroxide (797 mg,32.9 mmol) and water (20 mL). After stirring at RT for 5 h, adjust thepH of the reaction mixture to 5 with aqueous HCl (1M). Remove volatilesolvents in-vacuo to give an aqueous slurry. Filter and dry the solid toobtain the title compound (2.24 g, 88%). ES/MS m/z 381 (M+H).

Preparation 202-[4-[6-[(4-Cyano-2-fluoro-phenyl)methoxy]-2-pyridyl]-2,6-difluoro-phenyl]aceticacid

Prepare the title compound essentially as described in Preparation 18using methyl2-[4-[6-[(4-cyano-2-fluoro-phenyl)methoxy]-2-pyridyl]-2,6-difluoro-phenyl]acetate.ES/MS m/z 399 (M+H).

Preparation 212-[4-[6-[(4-Cyano-2-fluoro-phenyl)methoxy]-2-pyridyl]-2-fluoro-3-methyl-phenyl]aceticacid

Prepare the title compound essentially as described in Preparation 17using methyl2-[4-[6-[(4-cyano-2-fluoro-phenyl)methoxy]-2-pyridyl]-2-fluoro-3-methyl-phenyl]acetate.ES/MS m/z 395 (M+H).

Preparation 222-[4-[6-[(4-Cyano-2-fluoro-phenyl)methoxy]-2-pyridyl]phenyl]acetic acid

Mix together 4-[(6-bromo-2-pyridyl)oxymethyl]-3-fluoro-benzonitrile(0.70 g, 2.3 mmol) and 2-(4-boronophenyl)acetic acid (0.64 g, 3.4 mmol),THF (15 mL), water (5 mL) and potassium carbonate (0.63 g, 4.6 mmol).Purge the mixture with nitrogen for 10 min, then add Pd(dppf)Cl₂ (0.085g, 0.11 mmol) and heat the mixture at 75° C. for 8 h. Acidify themixture to pH 4-5 with aqueous HCl (1 M). Partition the mixture betweenEtOAc (50 mL) and water (50 mL). Wash the organics with brine (50 mL),dry over (Na₂SO₄), then filter and concentrate. Purify the residue bysilica gel chromatography using a gradient of 25 to 65% EtOAc in hexanesto give the title compound (800 mg, 97% yield) as solid. ES/MS m/z 363.0(M+H).

Preparation 23 Methyl 4-amino-3-[[(2S)-oxetan-2-ylmethyl]amino]benzoate

To a solution of methyl 3-fluoro-4-nitro-benzoate (2.0 g, 10 mmol) inTHF (10 mL) and DMF (10 mL) add triethylamine (3.1 mL, 22 mmol) at RT.To the slightly yellow solution add [(2S)-oxetan-2-yl]methanamine(Austin Chemical Company, 1.0 g, 11 mmol) and stir the rust-coloredsolution overnight. Dilute the reaction with EtOAc (100 mL) and water(50 mL). Separate the organic layer and then back-extract the aqueouslayer with EtOAc (2×50 mL). Combine the organics and wash with saturatedaqueous NaCl. Dry the organics over Na₂SO₄, filter, concentrate, and drythe residue under high vacuum. This gives crude methyl4-nitro-3-[[(2S)-oxetan-2-ylmethyl]amino]benzoate (2.8 g, 10 mmol) as ayellow solid (ES/MS m/z 267 (M+H)).

Next, dissolve methyl 4-nitro-3-[[(2S)-oxetan-2-ylmethyl]amino]benzoate(2.8 g, 10 mmol) in THF (50 mL) and add palladium on carbon (5%pre-wetted with water, 0.5 g). Vacuum purge the reaction mixture withhydrogen then stir under a balloon of hydrogen at RT for 2 h, duringwhich time the yellow color vanishes. Filter the mixture through Celite®and concentrate to give the title compound (2.4 g, 99%). ES/MS m/z 237(M+H).

Preparation 24 Methyl4-[[2-[4-[6-[(4-cyano-2-fluoro-phenyl)methoxy]-2-pyridyl]-3-methyl-phenyl]acetyl]amino]-3-[[(2S)-oxetan-2-ylmethyl]amino]benzoate

To a vial add2-[4-[6-[(4-cyano-2-fluoro-phenyl)methoxy]-2-pyridyl]-3-methyl-phenyl]aceticacid (1.10 g, 2.92 mmol), DMF (10 mL), HATU (1.4 g, 3.6 mmol), methyl4-amino-3-[[(2S)-oxetan-2-ylmethyl]amino]benzoate (0.76 g, 3.2 mmol),and DIPEA (1.5 mL, 8.6 mmol). Stir the mixture at RT for 30 min, thenpartition between EtOAc (30 mL) and water (30 mL). Wash the organicswith saturated aqueous NaCl (30 mL), then dry over Na₂SO₄, filter, andconcentrate. Purify the residue by silica gel chromatography using agradient of 10 to 35% EtOAc in DCM to give the title compound (1.2 g,69%) as solid. ES/MS m/z 595 (M+1), 593 (M−1).

Preparation 25 Methyl4-[[2-[4-[6-[(4-cyano-2-fluoro-phenyl)methoxy]-2-pyridyl]-2-fluoro-5-methyl-phenyl]acetyl]amino]-3-[[(2S)-oxetan-2-ylmethyl]amino]benzoate

To a flask add:2-[4-[6-[(4-cyano-2-fluoro-phenyl)methoxy]-2-pyridyl]-2-fluoro-5-methyl-phenyl]aceticacid (1.20 g, 3.04 mmol), DMF (15 mL), HATU (1.2 g, 3.1 mmol), methyl4-amino-3-[[(2S)-oxetan-2-ylmethyl]amino]benzoate (0.80 g, 3.4 mmol) andDIPEA (1.5 mL, 8.6 mmol). Stir the mixture at RT for 30 min, thenpartition between EtOAc (30 mL) and water (30 mL). Wash the organicswith saturated aqueous NaCl (30 mL), then dry over Na₂SO₄, filter andconcentrate. Purify the residue by silica gel chromatography using agradient of 10 to 35% EtOAc in DCM to give the title compound as solid(1.20 g, 64%). ES/MS m/z 613 (M+1), 611 (M−H).

Preparation 26 Methyl2-[[4-[6-[(4-cyano-2-fluoro-phenyl)methoxy]-2-pyridyl]-2-fluoro-phenyl]methyl]-3-[[(2S)-oxetan-2-yl]methyl]benzimidazole-5-carboxylate

To a round-bottom flask add2-[4-[6-[(4-cyano-2-fluoro-phenyl)methoxy]-2-pyridyl]-2-fluoro-phenyl]aceticacid (205 mg, 0.540 mmol), methyl4-amino-3-[[(2S)-oxetan-2-ylmethyl]amino]benzoate (116 mg, 0.490 mmol),HATU (224 mg, 0.589 mmol), DIPEA (0.26 mL, 1.5 mmol), and DMF (5 mL).After stirring at RT for 3.5 h, dilute the reaction mixture with EtOAc(30 mL) and wash with water and saturated aqueous NaCl. Dry the organicsover Na₂SO₄, filter, and concentrate. Purify the residue by silica gelchromatography using a gradient of 0 to 10% MeOH in DCM to obtain theintermediate amide (326 mg). ES/MS m/z 599 (M+H).

Heat the intermediate amide with acetic acid (5 mL) at 50° C. for 15 h.Concentrate the reaction mixture in-vacuo and dissolve the remainingresidue in EtOAc (25 mL). Wash the organics with saturated aqueousNaHCO₃ and saturated aqueous NaCl. Dry the organics over Na₂SO₄,filtered and concentrate. Purify the resulting by silica gelchromatography using a gradient of 20 to 100% EtOAc in hexanes to obtainthe title compound (152 mg, 52%). ES/MS m/z 581 (M+H).

Preparation 27 Methyl4-[[2-[4-[6-[(4-cyano-2-fluoro-phenyl)methoxy]-2-pyridyl]-2,6-difluoro-phenyl]acetyl]amino]-3-[[(2S)-oxetan-2-ylmethyl]amino]benzoate

Prepare the title compound essentially as described in Preparation 24using2-[4-[6-[(4-cyano-2-fluoro-phenyl)methoxy]-2-pyridyl]-2,6-difluoro-phenyl]aceticacid. Collect the product which precipitates during the aqueous workupby filtration and use without further purification. ES/MS m/z 617 (M+H),615 (M−H).

Preparation 28 Methyl4-[[2-[4-[6-[(4-cyano-2-fluoro-phenyl)methoxy]-2-pyridyl]-2-fluoro-3-methyl-phenyl]acetyl]amino]-3-[[(2S)-oxetan-2-ylmethyl]amino]benzoate

Prepare the title compound essentially as described in Preparation 24using 2-[4-15[6-[(4-cyano-2-fluoro-phenyl)methoxy]-2-pyridyl]-2-fluoro-3-methyl-phenyl]aceticacid. ES/MS m/z 613 (M+H), 611 (M−H).

Preparation 29 Methyl4-[[2-[4-[6-[(4-cyano-2-fluoro-phenyl)methoxy]-2-pyridyl]phenyl]acetyl]amino]-3-[[(2S)-oxetan-2-ylmethyl]amino]benzoate

Prepare the title compound essentially as described in Preparation 25using 2-[4-[6-[(4-cyano-2-fluoro-phenyl)methoxy]-2-pyridyl]phenyl]aceticacid. ES/MS m/z 581.0 (M+H), 579.0 (M−H).

Example 12-[[4-[6-[(4-Cyano-2-fluoro-phenyl)methoxy]-2-pyridyl]-3-methyl-phenyl]methyl]-3-[[(2S)-oxetan-2-yl]methyl]benzimidazole-5-carboxylicacid

To a vial add methyl4-[[2-[4-[6-[(4-cyano-2-fluoro-phenyl)methoxy]-2-pyridyl]-3-methyl-phenyl]acetyl]amino]-3-[[(2S)-oxetan-2-ylmethyl]amino]benzoate(1.2 g, 2.0 mmol) and acetic acid (6 mL). Heat the mixture at 80° C. for2 h, then remove the solvent in-vacuo. Partition the residue betweenEtOAc (30 mL) and aqueous NaHCO₃(5%, 20 mL). Wash the organics withsaturated aqueous NaCl (30 mL), dry over Na₂SO₄, filter, andconcentrate. Dissolve the residue in ACN (5 mL) and water (3 mL), thenadd to the mixture LiOH (0.22 g, 9.2 mmol) and stir at 50° C. for 2 h.Remove the solvent in-vacuo. Purify the residue by reverse-phase flashchromatography using a gradient of 20 to 35% ACN in 5% aqueous NH₄HCO₃to give the title compound (900 mg, 79%) as a solid. ES/MS m/z 563(M+H), 561 (M−H).

Example 22-[[4-[6-[(4-Cyano-2-fluoro-phenyl)methoxy]-2-pyridyl]-2-fluoro-5-methyl-phenyl]methyl]-3-[[(2S)-oxetan-2-yl]methyl]benzimidazole-5-carboxylicacid

To a vial add methyl4-[[2-[4-[6-[(4-cyano-2-fluoro-phenyl)methoxy]-2-pyridyl]-2-fluoro-5-methyl-phenyl]acetyl]amino]-3-[[(2S)-oxetan-2-ylmethyl]amino]benzoate(1.20 g, 1.96 mmol) and acetic acid (15 mL), then heat the mixture at80° C. for 2 h. Remove the solvent in-vacuo. Partition the residuebetween EtOAc (30 mL) and aqueous NaHCO₃(5%, 20 mL). Wash the organicswith saturated aqueous NaCl (30 mL), dry over Na₂SO₄, filter, andconcentrate. Dissolve the residue in ACN (10 mL) and water (4 mL), thenadd to the mixture LiOH (0.24 g, 10 mmol) and stir at 50° C. for 2 h.Acidify the mixture with saturated aqueous citric acid to pH=4-5. Removethe solvent in-vacuo. Purify the residue by reverse-phase flashchromatography using a gradient of 20 to 35% ACN in 5% aqueous NH₄HCO₃to give the title compound (745 mg, 66%) as a solid. ES/MS m/z 581(M+H), 579 (M−H).

Example 2atert-Butylammonium;2-[[4-[6-[(4-cyano-2-fluoro-phenyl)methoxy]-2-pyridyl]-2-fluoro-5-methyl-phenyl]methyl]-3-[[(2S)-oxetan-2-yl]methyl]benzimidazole-5-carboxylate

Method 1—Preparation without Seed Crystals

Suspend2-[[4-[6-[(4-cyano-2-fluoro-phenyl)methoxy]-2-pyridyl]-2-fluoro-5-methyl-phenyl]methyl]-3-[[(2S)-oxetan-2-yl]methyl]benzimidazole-5-carboxylicacid (555 mg, 0.96 mmol) in acetone (6 mL) while stirring at 800 rpm at50° C., giving a slurry of white solid. Add tert-butylamine (115 μL,1.09 mmol, 1.14 eq) observing a brief clarification of the mixturefollowed by precipitation of a white solid. Stir this slurry at 50° C.for 1 h, then turn off heating and allow the sample to stir as it comesto RT. Filter off the solid by vacuum filtration and dry in place for 15min under a stream of nitrogen, then dry in-vacuo at 50° C. for 1 h togive the title compound (612 mg, 98%).

Method 2—Preparation with Seed Crystals

Mix together2-[[4-[6-[(4-cyano-2-fluoro-phenyl)methoxy]-2-pyridyl]-2-fluoro-5-methyl-phenyl]methyl]-3-[[(2S)-oxetan-2-yl]methyl]benzimidazole-5-carboxylicacid (50 g, 86.1 mmol), acetone (658 mL), and water (42 mL), and heatthe mixture 50° C. Filter the mixture over GF/F paper and rinse with94:6 v:v acetone:water (25 mL). Heat the resulting solution at 50° C.Prepare a solution of tert-butylamine (10 mL, 94.7 mmol, 1.1 eq) and94:6 v:v acetone:water (25 mL). Add a portion of the tert-butylaminesolution (7 mL) followed by seed crystals oftert-butylammonium;2-[[4-[6-[(4-cyano-2-fluoro-phenyl)methoxy]-2-pyridyl]-2-fluoro-5-methyl-phenyl]methyl]-3-[[(2S)-oxetan-2-yl]methyl]benzimidazole-5-carboxylate(50 mg). Add the remaining tert-butylamine solution over approximately 1h via syringe pump at a rate of 0.47 mL/min. Heat the resultingsuspension at 50° C. for 2 h, then cool the mixture to ambienttemperature overnight. Filter the slurry and rinse with acetone (2×100mL). Dry the wetcake at 50° C. in-vacuo to a constant weight to give thetitle compound (51.8 g, 92%) as a pale yellow solid.

A prepared sample of the title compound is characterized by an XRDpattern using CuKα radiation as having diffraction peaks (2-thetavalues) as described in Table 1 below, and in particular having peaks at6.9 in combination with one or more of the peaks selected from the groupconsisting of 16.3 and 22.5; with a tolerance for the diffraction anglesof 0.2 degrees.

TABLE 1 X-ray powder diffraction peaks of tert-butylammonium;2-[[4-[6-[(4-cyano-2-fluoro-phenyl)methoxyl-2-pyridyl]-2-fluoro-5-methyl-phenyl]methyl]-3-[[(2S)-oxetan-2-yl]methyl]benzimidazole-5-carboxylate Angle (°2-Theta) RelativeIntensity Peak +/−0.2° (% of most intense peak) 1 5.5 26.20% 2 6.964.90% 3 11.2 49.20% 4 16.3 100.00%  5 17.1 34.70% 6 19.6 53.00% 7 21.843.10% 8 22.5 93.80% 9 27.3 41.10% 10 28.0 37.90%

Example 32-[[4-[6-[(4-Cyano-2-fluoro-phenyl)methoxy]-2-pyridyl]-2-fluoro-phenyl]methyl]-3-[(2S)-oxetan-2-ylmethyl]benzimidazole-5-carboxylicacid

Dissolve methyl2-[[4-[6-[(4-cyano-2-fluoro-phenyl)methoxy]-2-pyridyl]-2-fluoro-phenyl]methyl]-3-[[(2S)-oxetan-2-yl]methyl]benzimidazole-5-carboxylate(152 mg, 0.256 mmol) in THF (6 mL), then add lithium hydroxide (31 mg,1.26 mmol) and water (2 mL). Stir the mixture at RT for 16 h, thenadjust the pH to 6 with aqueous HCl (1N). Remove the THF in-vacuo andcollect the remaining solid by filtration. Purify by reverse-phase flashchromatography using a gradient of 10 to 40% ACN in aqueous NH₄HCO₃ (10mM, pH 10) to obtain the title compound (60 mg, 41%). ES/MS m/z 567(M+H).

Example 42-[[4-[6-[(4-Cyano-2-fluoro-phenyl)methoxy]-2-pyridyl]-2,6-difluoro-phenyl]methyl]-3-[(2S)-oxetan-2-ylmethyl]benzimidazole-5-carboxylicacid

Prepare the title compound essentially as described in Example 1 usingmethyl4-[[2-[4-[6-[(4-cyano-2-fluoro-phenyl)methoxy]-2-pyridyl]-2,6-difluoro-phenyl]acetyl]amino]-3-[[(2S)-oxetan-2-ylmethyl]amino]benzoate.Purify the product by reverse-phase flash chromatography using agradient of 30 to 50% ACN in aqueous NH₄HCO₃ (10 mM, pH 10). ES/MS m/z585 (M+H), 583 (M−H).

Example 52-[[4-[6-[(4-Cyano-2-fluoro-phenyl)methoxy]-2-pyridyl]-2-fluoro-3-methyl-phenyl]methyl]-3-[[(2S)-oxetan-2-yl]methyl]benzimidazole-5-carboxylicacid

Prepare the title compound essentially as described in Example 2 usingmethyl4-[[2-[4-[6-[(4-cyano-2-fluoro-phenyl)methoxy]-2-pyridyl]-2-fluoro-3-methyl-phenyl]acetyl]amino]-3-[[(2S)-oxetan-2-ylmethyl]amino]benzoate.Purify the product by reverse-phase flash chromatography using agradient of 5 to 40% ACN in 5% aqueous NH₄HCO₃. ES/MS m/z 581 (M+H), 579(M−H).

Example 62-[[4-[6-[(4-Cyano-2-fluoro-phenyl)methoxy]-2-pyridyl]phenyl]methyl]-3-[[(2S)-oxetan-2-yl]methyl]benzimidazole-5-carboxylicacid

Prepare the title compound essentially as described in Example 1 usingmethyl4-[[2-[4-[6-[(4-cyano-2-fluoro-phenyl)methoxy]-2-pyridyl]phenyl]acetyl]amino]-3-[[(2S)-oxetan-2-ylmethyl]amino]benzoate.Purify the crude product by reverse-phase flash chromatography using agradient of 20 to 35% ACN in 5% aqueous NH₄HCO₃. ES/MS m/z 549.0 (M+H),547.1 (M−H).

Biological Assays

Human GLP-1 Receptor HEK293 Cell cAMP Assay

GLP-1 Receptor functional activity is determined using cAMP formation inan HEK293 clonal cell line expressing human GLP-1R (NCBI accessionnumber NP_002053) at an expression density of 581±94 (n=6) and 104±12(n=5) fmol/mg protein (determined using [¹²⁵I]GLP-1(7-36)NH₂ homologouscompetition binding analysis). The hGLP-1R receptor expressing cells aretreated with compound (20 point concentration-response curve in DMSO,2.75-fold Labcyte Echo direct dilution, 384 well plate Corning Cat#3570) in DMEM (Gibco Cat #31053) supplemented with 1×GlutaMAX™ (GibcoCat #35050), 0.1% bovine casein (Sigma C4765-10ML), 250 IBMX(3-Isobutyl-1-methylxanthine, Acros Cat #228420010) and 20 mM HEPES(Gibco Cat #15630) in a 20 μL assay volume (final DMSO concentration is0.5%). After a 30 min incubation at 37° C., the resulting increase inintracellular cAMP is quantitatively determined using the CisBio cAMPDynamic 2 HTRF Assay Kit (62AM4PEJ). Briefly, cAMP levels within thecell are detected by adding the cAMP-d2 conjugate in cell lysis buffer(10 μL) followed by the antibody anti-cAMP-Eu³⁺-Cryptate, also in celllysis buffer (10 The resulting competitive assay is incubated for atleast 60 min at RT, then detected using a PerkinElmer Envision®instrument with excitation at 320 nm and emission at 665 nm and 620 nm.Envision units (emission at 665 nm/620 nm*10,000) are inverselyproportional to the amount of cAMP present and are converted to nM cAMPper well using a cAMP standard curve. The amount of cAMP generated (nM)in each well is converted to a percent of the maximal response observedwith human GLP-1(7-36)NH₂. A relative EC₅₀ value and percent top(E_(max)) are derived by non-linear regression analysis using thepercent maximal response vs. the concentration of compound added, fittedto a four-parameter logistic equation. The EC₅₀ and E_(max) data whenthe compounds of Examples 1-6 are tested in the cAMP assay describedabove using HEK293 cells expressing 581 and 104 fmol/mg GLP-1R are shownin Tables 2 and 3, respectively. These data indicate that the compoundsof Examples 1-6 are agonists of the human GLP-1 receptor.

TABLE 2 HEK293 cell line with 581 fmol/mg expression density of GLP-1R,intracellular cAMP response Example EC₅₀ (nM) ± SEM (n) E_(max) (%) ±SEM (n) 1  9.33 ± 1.36 (n = 6) 99.5 ± 2.53 (n = 6) 2 1.14 ± 0.315 (n =6)  104 ± 4.35 (n = 6) 3 3.08 ± 0.379 (n = 5)   99 ± 3.69 (n = 5) 4 3.99± 0.378 (n = 3)   99.2 ± 4 (n = 3) 5 6.45 ± 0.934 (n = 3)  105 ± 2.43 (n= 3) 6   20 ± 6.51 (n = 4)  101 ± 3.42 (n = 4)

TABLE 3 HEK293 cell line with 104 fmol/mg expression density of GLP-1R,intracellular cAMP response Example EC₅₀ (nM) ± SEM (n) E_(max) (%) ±SEM (n) 1 20 ± 3.25 (n = 6) 71.4 ± 2.26 (n = 6) 2 3.97 ± 0.61 (n = 6)   79.2 ± 3.2 (n = 6) 3  10 ± 2.3 (n = 5) 81.7 ± 3.86 (n = 5) 4 9.59 ±2.36 (n = 3)    78.3 ± 5.1 (n = 3) 5 23.6 ± 5.43 (n = 3)   76.7 ± 3.88(n = 3) 6 47.7 ± 17.9 (n = 4)    80.3 ± 3.1 (n = 4)

In Vivo Intraperitoneal Glucose Tolerance Test in Human GLP-1R Knock-InMice

The potency of the exemplified compounds to lower the concentration ofblood glucose in vivo is determined using mice expressing the humanGLP-1R (NCBI accession number NP_002053) from the mouse Glp-1r geneticlocus (Jun, L. S., et al., PLoS One. 2014 9:e93746). Overnight fastedmice are orally administered the test compound solubilized in10%Kolliphor® (HS15) in Polyetheylene Glycol 400 (PEG400). One hourpost-dose, the animals are administered glucose by intraperitonealinjection (2 g/kg), and blood glucose levels are measured intermittinglyover the next two hours using glucometers. A dose range of the testcompound is delivered, and area under the curve calculations for eachdose group are determined and fit to a four-parameter logistic model forcalculating in vivo potency as an ED50 with a 95% confidence interval.When tested in the in vivo intraperitoneal glucose tolerance testdescribed above, the compounds of Examples 1-3 exhibit potency to lowerthe concentration of blood glucose in mice expressing the human GLP-1Rwith ED50 (and 95% confidence interval) values as shown in Table 4,which indicates that these compounds are orally available potent GLP-1Ragonists in mice.

TABLE 4 Blood glucose lowering efficacy in mice expressing human GLP-1RBlood glucose lowering 95% confidence Example ED₅₀ (mg/kg) interval 10.09 0.0301-0.2592 2 0.07 0.0246-0.1808 3 0.06 0.013-0.246

Non-Human Primate (NHP) Pharmacokinetics:

The test compound is administered to fasted male cynomolgus monkeysintravenously (IV) at 0.5 mg/kg (using a dose volume of 1 mL/kg). Serialblood samples are collected at 0.08, 0.25, 0.5, 1, 2, 4, 8, 12, and 24hours post dose for IV bolus. After treatment with an EDTA coagulant,plasma is obtained by centrifugation and stored at −70° C. untilanalysis by LC-MS/MS. Test article concentration is determined inplasma. Noncompartmental analysis is used to calculate plasma clearanceand steady-state volume of distribution. Table 5 shows thepharmacokinetic data on the compounds of Examples 1-3 in this assay.These data, in part, are used to inform human mechanistic PK projectionswhich suggest a human pharmacokinetic profile to support once dailydosing.

TABLE 5 Cynomolgous monkey pharmacokinetic data Plasma clearance Volumeof Example (mL/min/kg) distribution (L/kg) Vehicle* 1 13 1.2 A 2 11 1.1A 3 6 1.1 B *Vehicle A—5% DMSO and 95% (20% CAPTISOL ® (w/v) in water;Vehicle B—20% captisol (w/v) in water + 1 mole equivalent NaOH

Phosphodiesterase 10 (PDE10) Enzyme Activity Assay

To generate phosphodiesterase 10A1 (PDE10A1) protein, a full-lengthPDE10A1 clone corresponding to GenBank ID AAD32595.1 is cloned intopFastBac1 (Invitrogen). The PDE10A1 protein with a C-terminal FLAG-tagis expressed by baculoviral infection of insect cells and purified usinganti-FLAG M2-agarose (Sigma) and size exclusion chromatography on aSuperdex 200 column (GE Healthcare) and stored at −80° C. in smallaliquots (20 mM Tris-HCl, Ph 7.5, 150 mM NaCl, 10% Glycerol).

PDE10A1 enzyme activities are measured with a yittrium silicate basedscintillation proximity assay (SPA) that detects radioactive nucleotidemonophosphates but not cyclic monophosphates. The assay buffer iscomposed of 50 mM Tris-HCl pH 7.5, 8 mM MgCl₂, 3.4 mM EDTA, and 0.1% BSA(Sigma). Assays are conducted in 384 well plates (3706, Corning) in atotal volume of 50 μl: comprised of 24 μl PDE10A1 enzyme, 1 μl testcompound and 25 μl of cyclic nucleotide. Test compounds are diluted inpure DMSO using ten-point concentration response curves with a 3-folddilution factor and 1 μl is acoustically dispensed into assay platesusing the Echo555 (LabCyte). 24 μl PDE10A1 protein is incubated with 1μl compound for 30 min before the reaction is started by the addition of[8-3H]-cGMP substrate (6.5 Ci/mmol, Perkin Elmer). Final concentrationof components is 70 μM PDE10A1, 80 nM (³H -cGMP), and 2% DMSO in assaybuffer. Maximal compound concentration in the reaction mixture is 10 μM.Reactions are incubated for 60 min at RT before quenching and theaddition of 400 mg/per well SPA beads (RPNQ0150, Perkin Elmer). Beadbound radioactivity (product) is quantified 12 h later with a Microbetacounter (Perkin Elmer). Data is normalized to % inhibition and IC₅₀values are calculated using the 4 parameter logistic equation asdescribed (Campbell, R M.; Dymshitz, J.; Eastwood, B. J.; et al. “DataStandardization for Results Management.” In: Sittampalam, G S.;Grossman, A.; Brimacombe, K.; et al.; eds. Assay Guidance Manual.Bethesda (Md.): Eli Lilly & Company and the National Center forAdvancing Translational Sciences; 2004). Table 6 shows the activity ofthe compounds of Examples 1˜4 in this assay. These data show that thecompounds of Examples 1 to 4 have weak binding affinity to PDE10A, whichindicates a reduced toxicity risk.

Human hERG K⁺ Channel Affinity Radioligand Binding Assay

The affinity of compounds for the human hERG K⁺channel in transfectedHEK-293 cells is evaluated in a radioligand binding assay as describedherein. Cell membrane homogenates (about 40 μg protein) are incubatedfor 60 min at 22° C. with 3 nM [³H]dofetilide in the absence or presenceof the test compound in a buffer containing 50 mM Tris-HCl (pH 7.4), 10mM KCl and 1 mM MgCl₂. The assay is carried out in a 96-well plateformat with a volume of 200 containing a maximum of 1% DMSO from initialsolubilization of test compound. Following incubation, the samples arefiltered rapidly under vacuum through glass fiber filters (GF/B,Packard) presoaked with 0.3% PEI and rinsed several times with ice-cold50 mM Tris-HCl, 10 mM KCl and 1 mM MgCl₂ using a 96-sample cellharvester (Unifilter, Packard). The filters are dried and then countedfor radioactivity in a scintillation counter (Topcount, Packard) using ascintillation cocktail (Microscint 0, Packard). Table 7 shows theactivity of Examples 1-3 in this assay, expressed as a percentinhibition of the control radioligand specific binding. These data showthat the compounds of Examples 1 to 3 have weak hERG inhibitoryactivity, which indicates a reduced toxicity risk.

TABLE 7 Human hERG K⁺ channel affinity radioligand percent inhibitionPercent inhibition (%) at Example 100 μM compound concentration, n = 1 10 2 54 3 37

1-20. (canceled)
 21. A compound selected from the group consisting of:

and a pharmaceutically acceptable salt thereof.
 22. The compoundaccording to claim 21, wherein the compound is selected from the groupconsisting:

and a pharmaceutically acceptable salt thereof.
 23. The compoundaccording to claim 21, wherein the compound is

or a pharmaceutically acceptable salt thereof.
 24. The compoundaccording to claim 21, wherein the compound is:

or a pharmaceutically acceptable salt thereof.
 25. A pharmaceuticalcomposition comprising a compound, or a pharmaceutically acceptable saltthereof, according to claim 21 and at least one pharmaceuticallyacceptable carrier, diluent, or excipient.
 26. A method of treating typeII diabetes mellitus in a mammal comprising administering to the mammalan effective amount of a compound according to claim 21, or apharmaceutically acceptable salt thereof.
 27. The method according toclaim 26, wherein the compound is administered orally.
 28. A method oflowering blood glucose levels in a mammal comprising administering aneffective amount of a compound according to claim 21, or apharmaceutically acceptable salt thereof.
 29. The method according toclaim 28, wherein the compound is administered orally.
 30. A method oftreating hyperglycemia in a mammal comprising administering to themammal an effective amount of a compound according to claim 21, or apharmaceutically acceptable salt thereof.
 31. The method according toclaim 30, wherein the compound is administered orally.